Abstract
An efficient preparation of Periplaneta americana nymphae allergen, Cr PI (54 kDa) is described. It was expressed as a GST-tag fusion protein in Escherichia coli, strain BL21 (DE3). Expression of recombinant Cr PI (rCr PI), denaturation/ renaturation of the inclusion bodies and the effects of protein and L-arginine concentration on inclusion body aggregation were optimized. The fusion protein was purified by affinity chromatography and size exclusion chromatography, and Cr PI fusion protein was purified to > 95%. rCr PI bound strongly to IgE in the sera of individuals with cockroach allergies as shown by western blot and ELISA. Highly refolded and purified recombinant protein was obtained, providing a basis for the large-scale preparation of Cr PI allergen.
Keywords: Periplaneta americana, Cr PI allergen, high-level expression, inclusion bodies, immunological characterization
Protein & Peptide Letters
Title: Expression, Purification, and Immunological Characterization of Cr PI
Volume: 14 Issue: 9
Author(s): Hai-Qiang Wu, Zhi-Gang Liu, Pi-Xin Ran, Zhen-Wen Zhou and Bo Gao
Affiliation:
Keywords: Periplaneta americana, Cr PI allergen, high-level expression, inclusion bodies, immunological characterization
Abstract: An efficient preparation of Periplaneta americana nymphae allergen, Cr PI (54 kDa) is described. It was expressed as a GST-tag fusion protein in Escherichia coli, strain BL21 (DE3). Expression of recombinant Cr PI (rCr PI), denaturation/ renaturation of the inclusion bodies and the effects of protein and L-arginine concentration on inclusion body aggregation were optimized. The fusion protein was purified by affinity chromatography and size exclusion chromatography, and Cr PI fusion protein was purified to > 95%. rCr PI bound strongly to IgE in the sera of individuals with cockroach allergies as shown by western blot and ELISA. Highly refolded and purified recombinant protein was obtained, providing a basis for the large-scale preparation of Cr PI allergen.
Export Options
About this article
Cite this article as:
Wu Hai-Qiang, Liu Zhi-Gang, Ran Pi-Xin, Zhou Zhen-Wen and Gao Bo, Expression, Purification, and Immunological Characterization of Cr PI, Protein & Peptide Letters 2007; 14 (9) . https://dx.doi.org/10.2174/092986607782110266
DOI https://dx.doi.org/10.2174/092986607782110266 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Subclinical Vitamin B12 Deficiency and Heart Rate Variability Across Life Cycle
Current Aging Science Patent Selections
Recent Patents on Inflammation & Allergy Drug Discovery The Roles of Mitochondrial Dysfunction and Reactive Oxygen Species in Aging and Senescence
Current Molecular Medicine A Practical Green Synthesis and Biological Evaluation of Benzimidazoles Against Two Neglected Tropical Diseases: Chagas and Leishmaniasis
Current Medicinal Chemistry Mass Spectrometric Analysis of F2-Isoprostanes: Markers and Mediators in Human Disease
Current Pharmaceutical Analysis Toll Like Receptors in Copd and Periodontal Disease-A Mini Review
Immunology, Endocrine & Metabolic Agents in Medicinal Chemistry (Discontinued) Sensitization to Inhaled Allergens in Atopic Children in Jordan Capital, Amman: A Cross-sectional Study
Current Respiratory Medicine Reviews The Safety of Bisphosphonate Use in Premenopausal Women on Corticosteroids
Current Drug Safety Novel Functions and Binding Mechanisms of Carbohydrate-Binding Proteins Determined by Force Measurements
Current Protein & Peptide Science Editorial [Hot Topic: Pharmacogenetic and Pharmacogenomics (Guest Editors: A. LLerena and J. Licinio) ]
Current Drug Targets Pharmacometrics of Stilbenes: Seguing Towards the Clinic
Current Clinical Pharmacology Metallothionein as a Negative Regulator of Pulmonary Inflammation
Current Pharmaceutical Biotechnology Plasma Proteins Interaction with Curcumin Nanoparticles: Implications in Cancer Therapeutics
Current Drug Metabolism Cardiovascular Biology of Interleukin-6
Current Pharmaceutical Design Multi-Component Synthesis of 6-Alkoxy-2-Amino-3,5-Dicyanopyridines
Letters in Organic Chemistry Meet Our Editorial Board Member
Current Organic Synthesis Drug-Induced Aseptic Meningitis
Current Drug Targets - Immune, Endocrine & Metabolic Disorders Mitogen-Activated Protein Kinases: New Molecular Targets for Pharmacological Treatment of Inflammatory Lung Diseases
Current Medicinal Chemistry - Anti-Inflammatory & Anti-Allergy Agents A Therapeutic Intervention With a Borderline Adolescent: The Inclusion in an Educational Community as a “Speaking Action”
Adolescent Psychiatry Evaluation and Management of Severe Asthma
Current Medicinal Chemistry